Collision-induced dissociation (CID), also known as collisionally activated dissociation (CAD), is a mass spectrometry technique to induce fragmentation of molecular ions in the gas phase. The molecular ions are usually accelerated by some electrical potential to high kinetic energy and then allowed to collide with neutral molecules (often helium, nitrogen or argon). In the collision some of the kinetic energy is converted into internal energy which results in bond breakage and the fragmentation of the molecular ion into smaller fragments. These fragment ions can then be analyzed by a tandem mass spectrometry.
Contents
- Triple quadrupole mass spectrometers
- Fourier transform ion cyclotron resonance
- Higher energy collisional dissociation
- Fragmentation mechanisms
- References
CID and the fragment ions produced by CID are used for several purposes. Partial or complete structural determination can be achieved. In some cases identity can be established based on previous knowledge without determining structure. Another use is in simply achieving more sensitive and specific detection. By detecting a unique fragment ion, the precursor ion can be detected in the presence of other ions of the mass to charge ratio, reducing the background and increasing the limit of detection.
Triple quadrupole mass spectrometers
In a triple quadrupole mass spectrometer there are three quadrupoles. The first quadrupole termed "Q1" can act as a mass filter and transmits a selected ion and accelerates it towards "Q2" which is termed a collision cell. The pressure in Q2 is higher and the ions collides with neutral gas in the collision cell and fragments by CID. The fragments are then accelerated out of the collision cell and enter Q3 which scans through the mass range, analyzing the resulting fragments (as they hit a detector). This produces a mass spectrum of the CID fragments from which structural information or identity can be gained. Many other experiments using CID on a triple quadrupole exist such as precursor ion scans that determines where a specific fragment came from rather than what fragments are produced by a given molecule.
Fourier transform ion cyclotron resonance
Sustained off-resonance irradiation collision-induced dissociation (SORI-CID) is a CID technique used in Fourier transform ion cyclotron resonance mass spectrometry which involves accelerating the ions in cyclotron motion (in a circle inside of an ion trap) and then increasing the pressure resulting collisions that produce CID fragments.
Higher-energy collisional dissociation
Higher-energy collisional dissociation (HCD) is a CID technique specific to the orbitrap mass spectrometer in which fragmentation takes place external to the trap. HCD was formerly known as higher-energy C-trap dissociation. In HCD, the ions pass through the C-trap and into the HCD cell, an added multipole collision cell, where dissociation takes place. The ions are then returned to the C-trap before injection into the orbitrap for mass analysis. HCD does not suffer from the low mass cutoff of resonant-excitation (CID) and therefore is useful for isobaric tag–based quantification as reporter ions can be observed. Despite the name, the collision energy of HCD is typically in the regime of low energy collision induced dissociation (less than 100 eV).
Fragmentation mechanisms
Homolytic fragmentation is bond dissociation where each of the fragments retains one of the originally-bonded electrons.
Heterolytic fragmentation is bond cleavage where the bonding electrons remain with only one of the fragment species.
In CID, charge remote fragmentation is a type of covalent bond breaking that occurs in a gas phase ion in which the cleaved bond is not adjacent to the location of the charge. This fragmentation can be observed using tandem mass spectrometry.